Abstract

TFIIB is an RNA polymerase II general transcription factor (GTF) that has also been implicated in the mechanism of action of certain promoter-specific activators (see, for examples, [1-11]). TFIIB enters the preinitiation complex (PIC) primarily through contact with the TATA box binding protein (TBP), an interaction mediated by three TBP residues [12-14]. To study the role of TFIIB in transcription activation in vivo, we randomly mutagenized these three residues in yeast TBP and screened for promoter-specific activation mutants. One mutant bearing a single conservative substitution, TBP-E186D, is the focus of this study. As expected, TBP-E186D binds normally to the TATA box but fails to support the entry of TFIIB into the PIC. Cells expressing TBP-E186D are viable but have a severe slow-growth phenotype. Whole-genome expression analysis indicates that transcription of 17% of yeast genes are compromised by this mutation. Chimeric promoter analysis indicates that the region of the gene that confers sensitivity to the TBP-E186D mutation is the UAS (upstream activating sequence), which contains the activator binding sites. Most interestingly, other TBP mutants that interfere with different interactions (TFIIB, TFIIA, or the TATA box) and a TFIIB mutant defective for interaction with TBP all manifest distinct and selective promoter-specific activation defects. Our results implicate the entry of TFIIB into the PIC as a critical step in the activation of certain promoters and reveal diverse mechanisms of transcription activation.

Interaction Annotations

Increase the total number of rows showing on this page by using the pull-down located below the table, or use the
page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column
header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small
"i" buttons located within a cell for an annotation to view further details about experiment type and any other
genes involved in the interaction.

Gene Ontology Annotations

Increase the total number of rows showing on this page using the pull-down located below the table, or use the page
scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header
to sort by that column; filter the table using the "Filter" box at the top of the table.

Phenotype Annotations

Increase the total number of rows showing on this page using the pull-down located below the table, or use the page
scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header
to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i"
buttons located within a cell for an annotation to view further details.

Regulation Annotations

Increase the total number of rows displayed on this page using the pull-down located below the table, or use the
page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column
header to sort by that column; to filter the table by a specific experiment type, type a keyword into the Filter box
(for example, “microarray”); download this table as a .txt file using the Download button or click Analyze to
further view and analyze the list of target genes using GO Term Finder, GO Slim Mapper, SPELL, or YeastMine.

Expression Datasets

Increase the total number of rows displayed on this page using the pull-down located below the table, or use the
page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column
header to sort by that column; filter the table using the "Filter" box at the top of the table; download this table
as a .txt file using the Download button;